Validation of the first step of the heuristic refinement method for the derivation of solution structures of proteins from NMR data

A new method for the analysis of NMR data in terms of the solution structure of proteins has been developed. The method consists of two steps: first a systematic search of the conformational space to define the region allowed by the initial set of experimental constraints, and second, the narrowing of this region by the introduction of additional constraints and optional refinement procedures. The search of the conformational space is guided by heuristics to make it computationally feasible. The method is therefore called the heuristic refinement method and is coded in an expert system called PROTEAN. The paper describes the validation of the first step of the method using an artificial NMR data set generated from the known crystal structure of sperm whale carbon monoxymyoglobin. It is shown that the initial search procedure yields a low-resolution structure of the myoglobin molecule, accurately reproducing its main topological features, and that the precision of the structure depends on the quality of the initial data set.     

Ion-Exchange Chromatography Separates Activities Synthesizing and Degrading Fructose 2,6-Bisphosphate from C3 and C4 Leaves but Not from Rat Liver

Fructose-6-phosphate,2-kinase and fructose-2,6-bisphosphatase were separated on the basis of charge from leaves of C₃ (spinach, lettuce, and pea) and C₄ (sorghum and amaranthus) plants but not from rat liver—a tissue known to contain a bifunctional enzyme with both activities. [2-³²P]Fructose 2,6-bisphosphate binding experiments also suggest that the major forms of these activities reside on different proteins in leaves.     

Immunoreactivity to the pancreatic polypeptide family in the nervous system of the adult human blood fluke,Schistosoma mansoni

Summary The presence and distribution of neuropeptides belonging to the pancreatic polypeptide family have been demonstrated by an indirect immunofluorescence technique in the nervous systems of adult male and female Schistosoma mansoni. Seven antisera of differing regional specificity to pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY) were employed on both whole-mount and cryostat-sectioned material. Positive immunoreactivity (IR) was obtained with all antisera except an N-terminally-directed antiserum to NPY. In the CNS, immunoreactivity was restricted to cell bodies and nerve fibres in the anterior ganglia, central commissure and dorsal and ventral nerve cords of both sexes, whereas, in the PNS, positive-IR was present in the plexuses innervating the subtegumental musculature and the oral and ventral suckers. Intense immunoreactivity was observed in a plexus of nerve fibres and cell bodies in the lining of the gynaecophoric canal and in fine nerve fibres innervating the dorsal tubercles of the male. In contrast, in the female, strong immunoreactivity was evident in nerve plexuses innervating the lining of the ovovitelline duct and in the wall of the ootype, but most notably in a cluster of cells in the region of Mehlis' gland. Results suggest that molecules with C-terminal homology to the PP-family are present in S. mansoni. These peptides would appear to be important regulatory molecules in the parasite's nervous system and may play a role in the control of egg production.     

Influence of Synchro-Mate-B and breed composition on estrous response and pregnancy rate in spring- and fall-bred Brahman crossbred beef cows

Estrous response, pregnancy rate, and the relationship between palpable corpora lutea (CL) and plasma progesterone concentrations were studied during the spring (n = 149) and fall (n = 146) breeding seasons using postpartum, crossbred beef cows consisting of 0-Brahman, 1 4 - Brahman , or 1 2 Brahman breeding. At the start of each breeding season, a jugular blood sample was collected for progesterone analysis; each cow was palpated for the presence of a CL and randomly alloted within breed to a non-treated control or Synchro-Mate-B (SMB) treatment. Fewer (P<0.01) fall than spring bred cows exhibited estrus within the first 21 d of breeding. Within 48 h of implant removal, 84 and 64% (spring and fall, respectively) of synchronized cows exhibited estrus. In both seasons, more SMB than control cows became pregnant by 7 d of breeding. Percentage of Brahman breeding did not influence estrous response or pregnancy rate. On the basis of 292 evaluations, palpation per rectum correctly assessed ovarian status 74% of the time. These data indicate that although season can influence response. SMB can be used effectively to synchronize Brahman crossbred cows. In addition, it was found that there is a close relationship between palpation per rectum and plasma concentrations of progesterone.     

Comparison of a competitive enzyme immunoassay kit and the infant mouse assay for detecting Escherichia coli heat-stable enterotoxin

A commercial competitive enzyme immunoassay kit, Escherichia coli ST EIA, was compared with the conventional infant mouse assay for sensitivity and specificity in detecting E. coli heat-stable enterotoxin. Thirty-one of 46 strains of E. coli tested were positive by both assays, while 15 strains were negative. The sensitivity of the ST EIA kit was up to 64-fold lower than the infant mouse assay.     

Supplemental chloralose anesthesia in morphine premedicated dogs

This study evaluated the cardiorespiratory stability of six dose-regulated, 12-hour, chloralose anesthetic maintenance protocols. Thirty mongrel dogs were premedicated with morphine sulfate (5mg/kg) and anesthetized with an induction dose of chloralose (80mg/kg). Fifteen animals were permitted to breathe spontaneously and 15 animals were ventilated mechanically to maintain a constant arterial pCO2 (40 +/- 5 mmHg). The spontaneously breathing dogs were separated into three groups in which animals (n = 5) were given different bolus doses of supplemental anesthetic. Initially the spontaneously breathing animals were hypoxemic, acidemic and hypercapnic. No consistent hemodynamic difference was noted among these groups. The mechanically ventilated animals were also divided into three groups that received varying doses of supplemental chloralose by constant infusion. Significant (p less than 0.01) myocardial depression was noted in the heavy-dosed animals by the third hour. Systolic pressure decreased 44%, pulse pressure decreased 37% and peak left ventricular dP/dt decreased 52%. All heavy-dosed animals expired before the eighth hour. Although these data suggest that morphine-premedicated, chloralose-anesthetized animals generally provide a stable cardiopulmonary model, high-dose chloralose supplementation depressed ventilation and produced a dose-dependent cardiotoxicity.     

Feeding and the control of volume within the tests of regular sea-urchins

The rigid, fluid filled, echinoid test presents problems of internal volume control when the vital functions of the animal involve the introduction of material, from outside the test, into the internal fluid systems. The volume of food in the guts of echinoids can be shown to vary considerably in the short term, to an extent which demands considerable capabilities for internal volume compensation. This is achieved by adjusting the quantity of fluid held within the gut so that, although the volume of food may vary widely, the total volume of the gut remains constant within narrow limits. The volume of gut fluid varies inversely with the quantity of food, and ingestion of food through the mouth is compensated by ejection of an equal volume of fluid from the anus. Defaecation is compensated by intake of fluid into the gut. On starvation, the gut becomes completely fluid filled and free from food.
The food of regular urchins is habitually compacted into regularly sized subpherical pellets covered with a tough mucus coating. The formation of coated pellets takes place in the buccal cavity and pharynx. The pellets remain discrete throughout their passage along the gut. The mucus coating resists digestion and remains intact even after defaecation. The functional morphology of the buccal cavity and pharynx is described in detail.
It is suggested that the functional significance of the pellets is associated with the gut fluid, volume compensation mechanism, which requires free flow of fluid throughout the length of the gut. It is also suggested that the siphon may be implicated in volume control, where fine adjustment of gut volume can be achieved by balancing ejection of fluid from the anus with intake of water through the mouth, but that the water intake is by the siphonal route, thus avoiding the necessity of flushing and diluting the enzyme rich digestive stomach region.